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首页> 外文期刊>Infection and immunity >A 140-kilodalton extracellular protein is essential for the accumulation of Staphylococcus epidermidis strains on surfaces.
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A 140-kilodalton extracellular protein is essential for the accumulation of Staphylococcus epidermidis strains on surfaces.

机译：140公斤的细胞外蛋白对于表皮葡萄球菌菌株在表面上的积累至关重要。

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Two distinct pathogenic mechanisms, adhesion to polymer surfaces and subsequent accumulation of sessile bacterial cells, are considered important pathogenic steps in foreign body infections caused by Staphylococcus epidermidis. By using mitomycin mutagenesis, we have recently generated a mutant, strain M7, from S. epidermidis RP62A which is unaffected in adhesion but deficient in accumulation on glass or polystyrene surfaces and lacks a 115-kDa extracellular protein (designated the 140-kDa antigen; F. Schumacher-Perdreau, C. Heilmann, G. Peters, F. G?tz, and G. Pulverer, FEMS Microbiol. Lett. 117:71-78, 1994). To evaluate the role of this protein in accumulation, we harvested extracellular proteins from S. epidermidis RP62A grown on dialysis membranes placed over chemically defined medium, purified the protein by using ion-exchange chromatography, determined its N-terminal amino acid sequence, and raised antiserum in rabbits. The antibody recognized only a single band in a Western immunoblot of the crude extracellular extract. With the microtiter biofilm test, antiserum at a dilution of < or =1:1,000 blocked accumulation of RP62A up to 98% whereas preimmune serum did not. The 140-kDa antigen was found only in extracellular products from bacteria grown under sessile conditions. Of 58 coagulase-negative clinical isolates, 32 strains were 140-kDa antigen positive and produced significantly larger amounts of biofilm than the 26 strains that were 140-kDa antigen negative. The 140-kDa protein appears to be biochemically and functionally unrelated to any previously described factors associated with biofilm formation. Thus, the 140-kDa antigen, referred to as accumulation-associated protein, may be a factor essential in S. epidermidis accumulation and, due to its immunogenicity, may allow the development of novel immunotherapeutic strategies for prevention of foreign body infection.
机译：在表皮葡萄球菌引起的异物感染中，两种不同的致病机制，即与聚合物表面的粘附和随后的无柄细菌细胞的积累，被认为是重要的致病步骤。通过使用丝裂霉素诱变，我们最近从表皮葡萄球菌RP62A产生了一个突变株M7，该菌株不粘附，但在玻璃或聚苯乙烯表面的积累不足，并且缺少115 kDa的细胞外蛋白（称为140 kDa抗原; F.Schumacher-Perdreau，C.Heilmann，G.Peters，F.G？tz和G.Pulverer，FEMS Microbiol.Lett.117：71-78，1994）。为了评估该蛋白质在积累中的作用，我们从生长在化学成分确定的培养基上的透析膜上的表皮葡萄球菌RP62A收获了细胞外蛋白质，通过离子交换色谱法纯化了该蛋白质，确定了其N端氨基酸序列，并进行了纯化。兔抗血清。该抗体在粗细胞外提取物的Western免疫印迹中仅识别一条带。用微量滴定生物膜测试，稀释度小于或等于1：1,000的抗血清可阻止RP62A积累高达98％，而免疫前血清则不能。仅在无梗条件下生长的细菌的细胞外产物中发现了140 kDa抗原。在58个凝固酶阴性临床分离株中，有32个菌株的140-kDa抗原呈阳性，与26个菌株的140-kDa抗原呈阴性相比，产生的生物膜量要大得多。 140-kDa蛋白似乎与任何先前描述的与生物膜形成有关的因素在生物化学和功能上均无关。因此，被称为积累相关蛋白的140 kDa抗原可能是表皮葡萄球菌积累中必不可少的因素，并且由于其免疫原性，可以开发出预防异物感染的新型免疫治疗策略。

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